IOP_Conf._Ser.__Earth_Environ._Sci._1289_012022

IOP Conference Series: Earth and Environmental Science
PAPER • OPEN ACCESS You may also like

- Effect of Adding Different Levels of
Leaf extract effectiveness of pecut kuda Vitamin D3 to Diet and its Relationship to
Qualitative Traits and Hatchability for
(Stachytarpheta jamaicensis L. Vahl) on Japanese Quail
M S Dawod and M A Al-Bayar
barramundi (Lates calcarifer) eggs hatchability - The Breeding of Free-range Chickens and
Purebred Chickens with Artificial
Insemination Technology on Fertility and
To cite this article: P Hartami et al 2023 IOP Conf. Ser.: Earth Environ. Sci. 1289 012022 Hatchability Using Manual Incubator
A. Tanjungsari, S. Andaruisworo and E.
Yuniati
- Effects of weight and storage duration of

hatching eggs of Indonesian Local
View the article online for updates and enhancements. Chicken on several measures of internal
quality and hatchability
D P Rahardja, M R Hakim, T Empra et al.
This content was downloaded from IP address 114.122.21.20 on 31/12/2023 at 01:15

5th International Symposium on Marine and Fisheries Research IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1289 (2023) 012022 doi:10.1088/1755-1315/1289/1/012022
Leaf extract effectiveness of pecut kuda (Stachytarpheta

jamaicensis L. Vahl) on barramundi (Lates calcarifer) eggs
hatchability
P Hartami1*, Mahdaliana1, Mainisa1, S Adhar1, M Hatta1, E Ayuzar1, DF
Hanum2, Rasidi3
1
Department of Aquaculture, Faculty of Agriculture, Malikussaleh University, Kampus
Utama Reuleut, Jl. Cot Tgk. Nie Muara Batu, Indonesia
2
Graduate Student of Aquaculture Department, Faculty of Agriculture, Malikussaleh
University, Kampus Utama Reuleut, Jl. Cot Tgk. Nie Muara Batu, Indonesia
3
Center for Fisheries Research, National Research and Innovation Agency, Gedung Biologi,
Jl. Raya Bogor KM. 47 Nanggewer Mekar Bogor West Java, Indonesia
Corresponding author mail: prama.hartami@unimal.ac.id
Abstract. Barramundi is one of the preferred fishes in great demand because of its delicious taste
and high nutritional content. One factor that influences the success of culture is hatchability and
high larval survival. In the business of barramundi aquaculture, problems are often encountered,
namely the presence of fungi that attack fish eggs. Precautions and treatment use of continuous
synthetic compounds with improper concentration will cause new problems. The traditional
pecut kuda anti-fungal, this plant is also easily obtainable and easy to use in the prevention and
treatment of fish diseases. The purpose of this study was to analyze the success rate of using
pecut kuda (Stachytarpheta jamaicensis L. Vahl) in increasing the hatchability of barramundi
eggs and observing the growth of barramundi larvae. This research was conducted on July 25 –
August 16, 2022. Located at Brackish Water Cultivation Fishery Center, Ujoeng Batee, Aceh.
This research method used a completely randomized design with 5 treatments and 3 replications,
namely P1 (0 ml/control), P2 (2 ml/L), P3 (4 ml/L), P4 (6 ml/L), and P5 (8 ml/L). Observation
of the data collected during the research was presented in the form of tables and graphs. The
results showed that pecut kuda leaf extract at different doses affected the hatchability of eggs,
percentage of fungus attack, and mortality of barramundi larvae, while it did not affect the
abnormality and growth length of barramundi larvae. The best treatment at P3 with a dose of 4
ml/L was 90.33% of the hatching rate with a low mortality value of P3 was 11.80%.
1. Introduction
Barramundi (Lates calcarifer) is one of the premium marine and brackish water aquaculture products
in Indonesia because of its relatively fast growth rate and ability to adapt to fluctuations in environmental
change [1], this makes barramundi suitable for small and large-scale aquaculture companies. It has been
proven that barramundi can be cultured in ponds or floating net cages in brackish water or seawater [2],
[3]. Barramundi also has market demand that continues to increase both domestically and internationally
[4]. In addition, barramundi is also one of the types of fish loved by many people because of its delicious
taste and high nutritional value. The successful development of barramundi aquaculture is largely
determined by the supply of good quality and quantity broodstock, eggs and larvae [6]. One of the factors
affecting the success of the farming crop is the high egg-hatching rate and larval survival rate, so
barramundi seeds are always available [1]. Obstacles to the availability of barramundi seed often include
the existence of fungi that attack fish eggs, both unfertilized and fertilized, leading to reduced
Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd 1
hatchability and survival rates. of larvae. [7]. The fungus that commonly attacks barramundi (Lates
calcarifer) eggs is Saproglenia sp, a fungal infection of eggs commonly known as fish mold [8]. Fungi
can attack eggs and reproduce there because of wounds caused by bacterial attack or mishandling [9].
According to [10], preventive and therapeutic measures often use synthetic compounds that have been
shown to have antifungal effects to improve egg quality. But on the other hand, continuing to use
chemicals and antibiotics at inappropriate concentrations will lead to new problems, including increased
resistance to fungi and negative impacts on the environment and humans.
The use of synthetic chemicals and antibiotics in large-scale farming is ineffective due to the
relatively high cost of antibiotics. Therefore, it is necessary to use alternative natural materials to treat
and prevent diseases, which are more effective and less expensive [12]. This alternative material is not
mold-resistant and is environmentally friendly. One of the medicinal plants with high properties that can
be used as a natural fungicide is pecut kuda leaves [13], [14]. These ingredients contain active
phytochemicals in the form of alkaloids, phenolic compounds, tannins, terpenoids, saponins and
flavonoids that act as antioxidants, bactericides, fungicides and anti-inflammatory agents [15]. There is
currently no information on the use of extracts of these natural ingredients to prevent and inhibit fungal
infections as well as increase the hatchability of barramundi (Lates calcarifer) eggs. Therefore, research
must be done to determine the ideal concentration of pecut kuda leaf in the incubation of barramundi
eggs and possibly provide a solution to combat the low production of barramundi seeds. This study aims
to test, analyze and evaluate the effectiveness of phytopharmaca raw material extract (pecut kuda leaves)
applied to the incubation environment with different doses on egg hatching and survival rate of
barramundi fry (Lates calcarifer).
2. Research method
2.1 Time and place
This experiment was done from August to September 2023 at the Brackish Water Cultivation Fisheries
Center in Ujong Batee Aceh Besar.
2.2 Research methodology

This research utilized a fully randomized design with five treatments and three replications. The dosage
used refers to research [16] for treatment using pecut kuda leaf without changing water up to 10 days of
rearing after hatching. The treatment used in detail was presented as follows.
P1 : Control (0 ml/L)
P2 : 2 ml/L
P3 : 4 ml/L
P4 : 6 ml/L
P5 : 8 ml/L
2.3 Research procedure

2.3.1 Extraction processing
The preparation of pecut kuda leaf extract was carried out at the Hatchery and Cultivation Technology
Laboratory, Faculty of Agriculture, University of Malikussaleh, using the maceration method. The
obtained leaves are first dried and then ground into powder and the dry weight is weighed as 500 grams
of powder. In addition, each ingredient is soaked with 96% ethanol at a ratio of 1: 5 at room temperature
20-250C, leave in a glass jar for 3 days and stirred well every day with a spoon. Additionally, evaporation
was performed at a temperature of 40–50°C to obtain a concentrated extract using a rotary evaporator
[17]. Once the extract was obtained, it was stored in a 500 ml glass container and then stored in the
freezer before testing at predetermined doses.
2.3.2 Container preparation

Before starting the study, the first step was to sterilize the equipment to be used by first washing it with
detergent and then drying it at room temperature. After drying, each plastic tub-shaped container with a
2
capacity of 2 litres for each treatment was filled with water using a filter, placed in the incubation room
and provided with an aeration system.
2.3.3 Preparation of barramundi eggs

The eggs used in this study were obtained from the spawning of barramundi at the Brackish Aquaculture
Fishery Center, Ujong Batee Aceh Besar, which looked clear and floated as a sign that 100 eggs had
been fertilized in the container. According to [18], eggs develop healthily, are transparent, and have a
bright colour so they are easy to distinguish from unfertilized eggs.
2.3.4 Soaking test

The extract of each ingredient was taken according to the dose of each treatment, put into each container,
diluted and added with water until each container reached a volume of 1 litre. Furthermore, each selected
egg was put into a test container for observation.
2.3.5 Larvae rearing

Larvae rearing during the study was carried out in 2 litres of incubation media with 1 litre of water
volume. Barramundi larvae were observed for 10 days, during maintenance no water changes and
siphoning were carried out because the larvae were still very vulnerable. Barramundi larvae 1-2 days
old were not given feed because there was still a yolk sack available in the larvae's body. Three-day-old
larvae were given rotifers until they were 10 days old at a density of 2-3 ind/ml every morning starting
at 08.00 WIB. According to [19], Rotifers were chosen as feed because they have good digestive
enzymes for larvae and complete nutritional content.
2.4 Observation parameters

2.4.1 Hatching rate
Hatching Rate (HR) was the hatchability of eggs or the number of eggs that hatch, the unit value of the
Hatching Rate was expressed in percent (%). For the calculation results of egg hatchability using the
formula [20] specifically.
୒୳୫ୠୣ୰୭୤୦ୟ୲ୡ୦ୣୢୣ୥୥ୱሺ୧୬ୢ୧୴୧ୢ୳ୟ୪ሻ
‫ܴܪ‬ሺΨሻ ൌ ͳͲͲ (1)
୘୭୲ୟ୪୬୳୫ୠୣ୰୭୤ୣ୥୥ୱ
2.4.2 Fungal infection percentage

To know the level of fungal infection in percent, prevalence calculations were carried out based on the
formula [21].
୒୳୫ୠୣ୰୭୤୧୬୤ୣୡ୲ୣୢୣ୥୥ୱ
ܲ‫݈݁ܿ݊݁ܽݒ݁ݎ‬ሺΨሻ ൌ ͳͲͲ (2)
୘୦ୣ୬୳୫ୠୣ୰୭୤ୣ୥୥ୱ୭ୠୱୣ୰୴ୣୢ
2.4.3 Larvae abnormality

Observation of larval abnormalities based on abnormalities or defects in head shape, body shape, and
tail shape. These physical abnormalities were documented in the form of pictures recorded and then
discussed descriptively. Calculations were made to determine the magnitude of the abnormality as stated
by [22], with the following formula.
୅ୠ୬୭୰୫ୟ୪୬୳୫ୠୣ୰୭୤୪ୟ୰୴ୟୣ
‫ݏ݁݅ݐ݈݅ܽ݉ݎ݋ܾ݊ܣ‬ሺΨሻ ൌ ୘୦ୣ୬୳୫ୠୣ୰୭୤୦ୟ୲ୡ୦ୣୢ୪ୟ୰୴ୟୣ ͳͲͲ (3)
2.4.4 Larval growth

Growth measurements were carried out at the beginning of rearing and the end of rearing larvae using a
micrometre on a microscope. Growth in length was calculated by the formula [23].
୍୬௅௧ି ௅௢
ሺΨሻ ൌ ୲
ͳͲͲ (4)
3
Information:
SGR : Specific Growth Rate (%
InLt : Average body length at the end of rearing (individual)
InLo : Average body length at the start of rearing (individual)
t : Rearing period (day)
2.4.5 Survival rate

Survival Rate (SR) was the comparison of fish survival at the beginning to the end of the study. SR
calculation was done by using the formula [24].
୒୲
ሺΨሻ ൌ ୒଴ ͳͲͲ (5)
Information:
SR : Survival Rate (%)
Nt : Number of larvae living in the final stage (individual)
N0 : Number of larvae living in the early stage (individual)
2.4.6 Water quality parameters

Physical and chemical water parameters observed at the time of hatching and rearing include salinity,
temperature, pH, dissolved oxygen, and ammonia [25]. Temperature measurements were taken every
morning using a thermometer, while measurements of dissolved oxygen, salinity, pH, and ammonia
were taken at the starting and the end of this study.
2.5 Data analysis

Observation data was presented in tables and graphs, and then several stages of testing were carried out,
namely the normality test and homogeneity test. If the data analyzed was normally distributed and
homogeneous, a parametric statistical test was carried out using one-way ANOVA. If the significance
value indicates a difference, then Tukey's further test was carried out to find out the difference in each
treatment.
3. Result
3.1 Hatching rate
The results showed that pecut kuda leaf extract with different doses was able to increase the hatchability
of barramundi fish eggs as shown in Figure 1 below.
100 90,33
84.33
90 79.67
80 73,33 71.00
Hatching rate (%)
70
60
50
40
30
20
10
0
P1 (0 ml/L) P2 (2 ml/L) P3 (4 ml/L) P4 (6 ml/L) P5 (8 ml/L)
Treatments
Figure 1. Graph of egg hatchability in each treatment.

It can be seen from Figure 1 that each treatment had different egg-hatching rates. The best hatching
rate was observed in the treatment P3 with a dose of 4 ml/L, which was 90.33%, while P5 with a dose
of 8 ml/L was the treatment with the lowest hatching value, namely 71%. Based on the ANOVA test, it
showed that administration of pecut kuda leaf extract had a significant effect on the hatchability of
4
barramundi fish eggs with a Fcount of 19.06 > Ftable (0.05) of 3.48. Tukey's further test results showed that
treatment P3 had a significant effect on other treatments, but was not significantly different from P2.
3.2 Fungus attack percentage

The results of the study of giving pecut kuda leaf extract to barramundi fish eggs showed the percentage
of fungal attacks as shown in Table 1 below.
Table 1. Percentage of fungal attack in each treatment.
Number of egg (grain)
Treatments Percentage (%)
Infected Checked
P1 (0 ml/L) 25 300 8.33
P2 (2 ml/L) 21 300 7.00
P3 (4 ml/L) 17 300 5.66
P4 (6 ml/L) 16 300 5.33
P5 (8 ml/L) 10 300 3.33
Table 1 describes the highest percentage of fungal attacks in treatment P1 (0 ml/L) with a value of
8.33%, and P5 (8 ml/L) is the treatment that has the lowest percentage of fungal attacks, namely 3.33%.
The following is a graphical picture of the level of resistance to fungal attack in each treatment.
100
96,67
Fungus attack resistance
98
94,33 94,67
96
93
94 91,67
92
(%)
90
88
86
Treatments
Figure 2. Fungus attack resistance level graph.

Figure 2 shows the highest level of fungal attack resistance in treatment P5 (8 ml/L) with a value of
96.67%, then P1 (0 ml/L) is the treatment that has the lowest fungal attack resistance level of 91.67%.
The results of the Anova test showed that administration of pecut kuda leaf extract had a significant
effect on the level of resistance to the fungal attack on barramundi eggs with a Fcount of 9.906 > Ftable
(0.05) of 3.48. Tukey's further test results showed that treatments P5, P4, and P3 had a significant effect
on other treatments. The type of fungus identified at the time of observation was Saprolegnia sp as
shown in Figure 3 below.
Figure 3. Saprolegnia sp which attacks barramundi fish eggs
5
The fungus that was found to be seen with the naked eye is white like cotton. After being observed
using a microscope, it can be seen that the fungus Saprolegnia sp. is long like threads and is grey-white
in colour.
3.3. Larvae abnormalities

The results of the study of giving pecut kuda leaf extract to barramundi eggs showed that there were no
abnormal barramundi larvae which can be seen in Figure 4 below.
Figure 4. Barramundi fish larvae

It can be seen in Figure 4, that the D10 age barramundi larvae looked normal for all treatments. In
the picture, it can be seen that the tail of the larvae is straight, the head size is not enlarged, the eyes are
normal size, and the colour of the white snapper larvae is also bright.
3.4. Growth length

The results showed that pecut kuda leaf extract at different doses did not affect the growth of barramundi
larvae as shown in Figure 5 below.
11
10,63
10,8 10.52 10.52 10,52
Growth length (%)
10,6
10,4
10,2 9,92
10
9,8
9,6
9,4
9,2
9
Treatments
Figure 5. Graph of specific growth rate for each treatment

It can be seen in Figure 5, that the highest percentage of specific growth rate during observation was
in treatment P4 (dose 6 ml/L) which was 10.63% per day, then P5 with a dose of 8 ml/L was a treatment
that had a specific growth rate percentage the lowest is 9.92% per day. Based on ANOVA, it showed
that administration of pecut kuda leaf extract did not affect the specific growth rate of barramundi with
Fcount 1.391 < Ftable (0.05) 3.48.
3.5. Larvae mortalities

The results showed that larval mortality had differences for each treatment as shown in Table 2, and the
survival rate is presented in Figure 6 below.
6
Table 2. Larvae mortality in each treatment.

Number of larvae (Individual)
Treatments Mortalities (%)
Hatching Dead
P1 (0 ml/L) 220 68 30.90
P2 (2 ml/L) 253 38 15.01
P3 (4 ml/L) 271 32 11.80
P4 (6 ml/L) 239 72 30.12
P5 (8 ml/L) 213 117 54.92
Table 2 shows that the highest mortality was in treatment P5 with a dose of 8 ml/L which was 54.92%
and P3 with a dose of 4 ml/L was the treatment that had the lowest mortality value, namely 11.80%. The
Survival Rate graph for each treatment can be seen in Figure 6.
100 84,98 88,19

Survival rate (%)
80 69,01 69,87
60 45,07
40
20
Treatments
Figure 6. Larvae survival rate.

As shown in Figure 6, the best survival rate was observed in treatment P3 with a dose of 4 ml/L
which was 88.19% and P5 with a dose of 8 ml/L was the treatment that had the lowest survival value,
namely 45.07%. Based on statistical analysis using the Anova test, it was shown that administration of
pecut kuda leaf extract had a significant effect on the survival rate of barramundi larvae with Fcount
43.004 > Ftable (0.05) 3.48. Tukey's further test results showed that the P2 and P3 treatments had a
significant effect on the other treatments.
3.6. Water quality parameters

The results of measurements of salinity, temperature, pH, and DO in research for egg-hatching media
and larval-rearing media are presented in the following table.
Table 3. Data on the measurement of water quality for hatching eggs
Average water quality data

Treatments
Temperatures (oC) pH Dissolved oxygen (ppm) Salinity (ppt)
P1 (0 ml/L) 27.00-27.80 7.80-8.00 5.00-5.50 28.00
P2 (2 ml/L) 27.80-27.90 7.60-8.00 4.67-5.30 28.00
P3 (4 ml/L) 27.50-28.00 7.80-8.10 4.28-5.20 28.00
P4 (6 ml/L) 27.40-27.90 7.90-8.10 4.27-5.21 28.00
P5 (8 ml/L) 27.80-28.10 7.90-8.20 4.29-5.20 28.00
7
Table 4. Data on the measurement of water quality for rearing eggs
Average water quality data

Treatments
Temperatures (oC) pH Dissolved oxygen (ppm) Salinity (ppt)
P1 (0 ml/L) 27.00-29.60 8.00-8.20 4.30-5.00 28.00-30.00
P2 (2 ml/L) 26.90-29.70 7.90-8.30 3.60-4.00 28.00-30.00
P3 (4 ml/L) 27.00-29.80 8.10-8.30 3.80-4.00 28.00-31.00
P4 (6 ml/L) 27.00-29.80 7.90-8.20 3.50-3.90 28.00-31.00
P5 (8 ml/L) 27.00-29.60 8.00-8.40 3.08-3.84 28.00-31.00
Tables 3 and 4 show that the water quality in the egg and larvae observation media is included in the
optimal category, namely temperature, pH, and salinity. Meanwhile, dissolved oxygen tends to decrease,
so it does not meet the optimal category for larval rearing even though it has been equipped with aeration
in each container.
4. Discussion
The highest hatching rate of barramundi eggs within 14 hours of observation in treatment P3 with a dose
of 4 ml/L was 90.33%. The value of the egg hatching rate obtained in this study almost coincides with
92.33% with the value of the hatching rate of eggs from research results [26] on the effectiveness of
using chemical disinfectants to Prevent fungal attacks on Labeo bata fish eggs. The dosage of 4 ml/L is
the dosage containing active ingredients such as alkaloids, flavonoids and other antifungal substances
that are completely optimal and suitable for barramundi, so at this dosage the eggs hatch well. The
content in pecut kuda leaves is not only an active antifungal agent but also contains some toxic
substances in large doses. Thus, at this dosage, the alkaloid active ingredient is an antifungal compound
that has good effects, while the tannic acid in the form of an acid that inhibits egg hatching has no
harmful effect because it is still at a low dosage.
Egg hatching is influenced by several factors, while egg failure can be caused by many factors such
as light intensity, temperature, pH, turbidity, dissolved oxygen content and ammonia. [27]. The 8 ml/L
dose contains more active ingredients so this concentration may affect the eggs. This is the opinion [28]
that hatching is not successful because the egg is highly permeable, so when fertilization begins, the egg
membrane absorbs water and the egg hatches quickly. If the incubation medium has a higher ion
concentration than the egg, the egg will be damaged because the liquid it contains will be absorbed by
the more concentrated medium. According to [29], in fertilized eggs, there is fluid that fills the space
around the spore, helping to provide space and protection for the developing embryo. The egg's
hatchability will decrease if the egg swells due to promoted osmosis. When the osmotic concentration
in the Perivitelline space is greater than the concentration of extracellular water surrounding the egg, the
extracellular water will move into the perivitelline space by osmosis, causing the egg to swell. Then, the
liquid inside the egg will flow out, the egg will shrink and die. Therefore, the P5 dose concentration is
not optimal for barramundi egg incubation.
The high hatching rate was also mainly determined by the low fungal attack rate in treatment P3,
which was 5.66%. The fungal attack percentage value in the extract treatment in this study was better
than the study by [30] who used clove leaf extract in African catfish hatchery at a dose of 11 ppm, or
50.77%. Through observation, the fungus found in this study is Saprolegnia sp with cotton fibre-like
characteristics, white to grey in colour. It is suggested [31] that Saprolegnia sp. fibrous, cotton-like,
white to grey and brown. The discovery of Saprolegnia sp during research supports the view [32] that
one of the fungi that attack barramundi eggs is Saprolegnia sp, commonly known as fish mold because
it attacks fish and eggs. fish. This fungus creates tangled excess mycelium, causing the living eggs
around the dead eggs to die. The fungus will interfere with the egg's respiration process and will
eventually die before hatching. There is an opinion [8] that when white barramundi eggs are attacked
by fungi, they will show signs of milky white eggs or smooth cotton-like fibres around the eggs.
8
Saprolegnia sp is characterized by the ability to move freely (have two flagella), the body's cells produce
filaments (hyphae do not have septa or branches), are filamentous and have no segments.
The ability of fungi to attack eggs decreased when increasing the concentration of pecut kuda leaf
extract used in the incubation media. This shows that the concentration of the extract affects fungal
attacks. According to [33], active compounds in the form of flavonoids, saponins, tannins, quinones,
steroid compounds, triterpenoids and alkaloids have good antifungal effects, so fish eggs are protected
from fungal attack because the fungal cells become sensitive to changes in the environment which causes
die. For P5 treatment, an extraction dose of 8 ml/L is the best dose to overcome fungal attack. This is
due to the content in pecut kuda leaves in the form of active alkaloids that protect the body's organs from
toxic substances, both in the form of parasites and fungi, as well as fungicidal antibacterial substances
has an inhibitory effect of some enzymes, leading to disturbances in the metabolism of fungal cells,
thereby inhibiting the elongation process of fungal hyphae. The fungal hyphae cannot be fragmented
because the mycelial tissue is damaged, so the fungal cells easily die [34]. However, the dose of P5 (8
ml/L) has a good inhibitory effect on fungi but does not inhibit egg hatching. The higher extract
concentration, not only has a more effective antifungal effect, but also kills barramundi eggs.
In addition to its ability to increase egg hatching rate and inhibit fungal attack, pecut kuda leaf extract
is also safe for barramundi larvae and therefore does not cause abnormalities during the hatching process
of the larvae. Indeed, pecut kuda leaf extract contains active compounds, including flavonoids. It has
been suggested [35] that flavonoid compounds can increase bone density, which is attributed to the
content of ethyl p-methoxycinnamate which belongs to the flavonoid group and is part of the
phytoestrogen group of compounds. The mechanism of action of phytoestrogen compounds in
regulating bone density may be similar to that of estrogen. Estrogen influences bone regeneration by
controlling the activity of osteoblasts and osteoclasts, including maintaining a balance in the activity of
the two cells by regulating the production of osteoclasts. paracrine factors [36]. Estrogen also affects
bone breakdown by inhibiting the maturation of osteoclasts, which in turn can inhibit bone resorption
(35).
Another benefit of pecut kuda leaf extract is that it does not harm the development of barramundi
larvae. Larval development in all treatments remained relatively normal, although there was a tendency
to decrease with increasing doses. The growth rate is influenced by competition for food and varying
levels of fish appetite. This is evident in statement [30] which states that growth can be influenced by
biotic and abiotic factors. Biological factors include age and the fish's ability to meet dietary
requirements, and abiotic factors include food availability and fish habitat quality. The highest SGR
found in this treatment was 10.63%, this value is higher than the results obtained in [30] on the effect of
soaked dose of clove leaf extract (Syzygium aromaticum) on catfish (Clarias gariepinus) egg hatching,
the best specific growth rate was achieved at the average of 0.252%.
Research results showed that pecut kuda leaf extract also had an impact on the low mortality rate of
barramundi larvae, especially in treatment P3, which was 11.80%. The highest mortality rate in this
study was 54.92%, this value is better than the results obtained by [37] regarding the effectiveness of
meniran plant extract (Phyllanthus niruri L.) against fungal attack (Saprolegnia sp.) on gouramy
(Osphronemus gouramy) egg hatchability, namely at a dose of 3 ml/l of 74.67%. The mortality rate of
larvae is estimated due to the administration of pecut kuda leaf extract which contains high tannin
compounds along with increasing doses. This follows the opinion [38] which states that there is an effect
of the tannin compounds contained in the leaf extract in the egg-hatching process, the tannin compounds
are acidic so that they can trigger the softening of the chorion layer which can cause the larvae to hatch
with a state of weakened immunity so they die easily. Thus, reducing fungal infection in eggs at high
doses that affect embryo viability will result in reduced egg hatchability.
Other factors affect the mortality rate of larvae, specifically the amount of dissolved oxygen during
rearing is relatively low, the optimal amount of dissolved oxygen for reared larvae is at least 4, while
the amount of dissolved oxygen obtained during observation is <4. This is consistent with the dissolved
oxygen value in seawater aquaculture, which must be greater than 4 mg/L [39]. According to [25], if the
amount of dissolved oxygen is unbalanced, the fish will be stressed because the brain is not provided
9
with enough oxygen and die due to lack of oxygen (anorexia) because the body does not receive enough
oxygen. [40] states that water quality is one of the factors that influence the production of aquaculture
activities, where the temperature parameter is a parameter that has a complex relationship with other
water qualities so that it can affect growth, reproduction, and survival.
The decrease in dissolved oxygen value was due to not changing the water during the research
process, so the ammonia concentration in the farming environment became high. It has been suggested
[41] that in aquaculture systems, for all water quality parameters, ammonia is the second limiting factor
after oxygen. At high concentrations, ammonia is toxic, significantly reducing oxygen supply and
causing unwanted changes in aquatic ecosystems. The water exchange or storage was not carried out
because the larvae were still too young, namely D1-D10. This is by the opinion [42], that early-stage
larvae from one to ten days old are sensitive to environmental factors as a result of water changes.
Environmental factors can be in the form of water quality parameters such as dissolved oxygen,
temperature, salinity, pH, and ammonia.
The temperature of the water media during egg rearing for all containers was stable and not too high,
namely around 27.00-28.10oC, and the temperature during larval rearing was around 26.90-29.60oC, this
corresponds to the temperature range for barramundi which is 27.00 -30.00ºC [43]. [44], added that egg
hatching will be faster at high temperatures because at high temperatures metabolic processes will occur
more quickly so that embryo development will also be faster and the movement of embryos in the shell
will be more intensive so that hatching will be faster. The pH of the media during the study for all
containers was in the range of 7.6-8.4 which is considered optimal, which is under the pH for barramundi
according to [39] that is, for the stages of rearing larvae in tubs and raising eggs are the same, ie, 7.00-
8.50. [29] said that a low pH can cause the work of the chorionase enzyme to be disrupted because this
enzyme cannot work optimally at a more acidic or lower pH.
5. Conclusion
The conclusion that can be drawn from this study was that administration of pecut kuda leaf extract
(Stachytarpheta jamaicensis L. Vahl) with different doses affects egg hatchability, percentage of fungal
attack, and larval mortality in barramundi (Lates calcarifer). The use of pecut kuda leaf extract can
reduce the egg mortality rate, thus giving a good value to the hatching rate. The use of pecut kuda leaf
extract with different tests did not affect the specific length growth rate of barramundi. The use of pecut
kuda leaf did not influence the advancement of barramundi larvae, so no abnormal larvae were found.
Providing pecut kuda leaf extract had no impact on changing the water quality of the barramundi egg or
larval culture environment.
References
[1] A. R. S. H. Saputra and S. H. Samara, “Performance analysis of white snapper (Lates calcarifer)
nursery at BBPBAP Jepara,” in IOP Conference Series: Earth and Environmental Science,
Institute of Physics, 2022. doi: 10.1088/1755-1315/1036/1/012118.
[2] D. S. Fielder, W. J. Bardsley, G. L. Allan, D. S. Fielder, and P. M. Pankhurst, “Effect of
photoperiod on growth and survival of snapper Pagrus auratus larvae,” Aquaculture, vol. 211,
no. 1–4, pp. 135–150, Aug. 2002, doi: 10.1016/S0044-8486(02)00006-6.
[3] N. M. Bacheler, K. W. Shertzer, B. J. Runde, P. J. Rudershausen, and J. A. Buckel,
“Environmental conditions, diel period, and fish size influence the horizontal and vertical
movements of red snapper,” Sci Rep, vol. 11, no. 1, Dec. 2021, doi: 10.1038/s41598-021-88806-
3.
[4] D. M. Cawthorn, C. Baillie, and S. Mariani, “Generic names and mislabeling conceal high
species diversity in global fisheries markets,” Conservation Letters, vol. 11, no. 5. Wiley-
Blackwell, Sep. 01, 2018. doi: 10.1111/conl.12573.
10
[5] Hardoko and S. Utami, “Chemical-physical properties characterization of white snapper fish skin
rambak crackers based on boiling and drying duration,” Jurnal Ilmiah Perikanan dan Kelautan,
vol. 12, no. 1, pp. 122–130, 2020, doi: 10.20473/jipk.v12i1.14842.
[6] Z. R. Paniza. “Broodstock management and seed production of marine fishes.” SEAFDEC Asian
Aquaculture, Vol. XXIII May-August 2001, no. 3-4.
[7] M. E. Bunker, G. Elliott, H. Heyer-Gray, M. O. Martin, A. E. Arnold, and S. L. Weiss,
“Vertically transmitted microbiome protects eggs from fungal infection and egg failure,” Anim
Microbiome, vol. 3, no. 1, Dec. 2021, doi: 10.1186/s42523-021-00104-5.
[8] F. Fadaeifard, M. Raissy, H. Bahrami, E. Rahimi, and A. Najafipoor, “Freshwater fungi isolated
from eggs and broodstocks with an emphasis on Saprolegnia in rainbow trout farms in west Iran,”
Afr J Microbiol Res, vol. 5, no. 22, Oct. 2011, doi: 10.5897/ajmr11.385.
[9] P. Kalatehjari, M. Yousefian, and M. A. Khalilzadeh, “Assessment of antifungal effects of
copper nanoparticles on the growth of the fungus Saprolegnia sp. on white fish (Rutilus frisii
kutum) eggs,” Egypt J Aquat Res, vol. 41, no. 4, pp. 303–306, Dec. 2015, doi:
10.1016/j.ejar.2015.07.004.
[10] R. Sun, J. Chen, C. Pan, Y. Sun, B. Mai, and Q. X. Li, “Antibiotics and food safety in
aquaculture,” J Agric Food Chem, vol. 68, no. 43, pp. 11908–11919, Oct. 2020, doi:
10.1021/acs.jafc.0c03996.
[11] X. Liu, J. C. Steele, and X. Z. Meng, “Usage, residue, and human health risk of antibiotics in
Chinese aquaculture: A review,” Environmental Pollution, vol. 223, pp. 161–169, 2017, doi:
10.1016/j.envpol.2017.01.003.
[12] Y. Fujaya, A. A. Hidayani, D. K. Sari, S. Aslamyah, N. Rukminasari, A. Muthalib, S. Cristianto,
E. Defista, H. Fazhan, and K. Waiho, “The optimal dosage of fermented herbal extract on growth
and feed efficiency of nile tilapia (Oreochromis niloticus).” Tropical Life Sciences Research, no.
1-22, November 2022.
[13] P. M. Liew, and Y. K. Yong, “Stachytarpheta jamaicensis (L.) Vahl: from traditional usage to
pharmacological evidence,” evidence-based complementary and alternative medicine, vol. 2016.
Hindawi Limited, 2016. doi: 10.1155/2016/7842340.
[14] P. D. Yadav, K. P. Modi, and M. B. Shah, “Phytochemistry, pharmacology, and botanical aspects
of Stachytarpheta species-A review,” International Journal of Green Pharmacy, vol. 15, no. 2,
p. 114, doi: 10.22377/ijgp.v15i2.3078.
[15] O. Zs, O. Oo, K. Se, and A. Oo, “Stachytarpheta jamaicensis leaf extract: Chemical composition,
antioxidant, anti-arthritic, anti-inflammatory and bactericidal potentials,” Journal of Scientific
and Innovative Research, vol. 6, no. 4, pp. 119–125, 2017, [Online]. Available:
www.jsirjournal.com
[16] L. A. Al_husnan and M. D. F. Alkahtani, “Impact of Moringa aqueous extract on pathogenic
bacteria and fungi in vitro,” Annals of Agricultural Sciences, vol. 61, no. 2, pp. 247–250, Dec.
2016, doi: 10.1016/j.aoas.2016.06.003.
[17] Q. W. Zhang, L. G. Lin, and W. C. Ye, “Techniques for extraction and isolation of natural
products: A comprehensive review,” Chinese Medicine (United Kingdom), vol. 13, no. 1.
BioMed Central Ltd., Apr. 17, 2018. doi: 10.1186/s13020-018-0177-x.
[18] M. A. Haque, M. I. Hossain, S. Aftabuddin, A. Habib, and M. A. M. Siddique, “First onboard
fertilization of Asian seabass, Lates calcarifer: Effects of egg stocking density on the
fertilization, hatching and survival rate,” Sci Afr, vol. 12, Jul. 2021, doi:
10.1016/j.sciaf.2021.e00841.
11
[19] M. Kailasam, A. R. Thirunavukkarasu, A. G. Ponniah, S. Selvaraj, and P. Stalin, “Recent

advances in rotifer culture and its application for larviculture of finfishes,” in Advances in Marine
and Brackishwater Aquaculture, Springer India, 2015, pp. 17–23. doi: 10.1007/978-81-322-
2271-2_2.
[20] M. A. Rahman, M. H. Rahman, S. M. Yeasmin, A. Al-Asif, and D. Mridha, “Identification of
causative agent for fungal infection and effect of disinfectants on hatching and survival rate of
bata (Labeo. Bata) larvae,” Advances in Plants & Agriculture Research, vol. 7, no. 4, Aug. 2017,
doi: 10.15406/apar.2017.07.00264.
[21] S. Elbashir, M. Jahncke, A. DePaola, J. Bowers, J. Schwarz, A. J. Punchihewage-Don, B. Min,
T. Rippen, and S. Parveen, “Prevalence and abundance of bacterial pathogens of concern in
shrimp, catfish and tilapia obtained at retail stores in Maryland, USA,” Pathogens, vol. 12, no.
2, Feb. 2023, doi: 10.3390/pathogens12020187.
[22] E. Shuba, D. Adriaens, and P. Bossier, “Variations in body shape of European seabass larvae
(Dicentrarchus labrax) reared under xenic and axenic conditions,” Ethiopian Journal of Science
and Technology, vol. 7, no. 2, p. 85, Jul. 2015, doi: 10.4314/ejst.v7i2.2.
[23] A. E. Sallam, M. M. M. El-feky, M. S. Ahmed, and A. T. Mansour, “Potential use of whey
protein as a partial substitute of fishmeal on growth performance, non-specific immunity and gut
histological status of juvenile European seabass, Dicentrarchus labrax,” Aquac Res, vol. 53, no.
4, pp. 1527–1541, Mar. 2022, doi: 10.1111/are.15688.
[24] A. T. Mukti, O. Carman, A. Alimuddin, M. Zairin, and M. A. Suprayudi, “Growth performance,
survival rate, flesh, and proximate composition of sex-grouped triploid and diploid Nile tilapia
(Oreochromis niloticus),” Turk J Vet Anim Sci, vol. 44, pp. 290–298, 2020, doi: 10.3906/vet-
1905-79.
[25] A. Bhatnagar, and P. Devi, “Water quality guidelines for the management of pond fish culture,”
Int J Environ Sci, vol. 3, no. 6, pp. 1980–2009, 2013, doi: 10.6088/ijes.2013030600019.
[26] M. A. Rahman, M. H. Rahman, S. M. Yeasmin, A. Al-Asif, and D. Mridha, “Identification of
causative agent for fungal infection and effect of disinfectants on hatching and survival rate of
bata (Labeo bata) Larvae,” Advances in Plants & Agriculture Research, vol. 7, no. 4, Aug. 2017,
doi: 10.15406/apar.2017.07.00264.
[27] M. Gomes, A. Correia, L. Pinto, C. Sá, V. Brotas, and M. Mateus, “Coastal water quality in an
atlantic sea bass farm site (Sines, Portugal): a first assessment,” Front Mar Sci, vol. 7, Apr. 2020,
doi: 10.3389/fmars.2020.00175.
[28] T. Myosho, H. Takahashi, K. Yoshida, T. Sato, S. Hamaguchi, T. Sakamoto, and M. Sakaizumi,
“Hyperosmotic tolerance of adult fish and early embryos are determined by discrete, single loci
in the genus Oryzias,” Sci Rep, vol. 8, no. 1, Dec. 2018, doi: 10.1038/s41598-018-24621-7.
[29] M. Kasiri, M. Sudagar, and S. A. Hosseini, “Effect of water hardness on egg hatchability and
larval viability of angelfish (Pterophyllum scalare (Schultze, 1823),” 2011. [Online]. Available:
http://www.urpjournals.com
[30] B. N. D. Yulihastiana, N. Cokrowati, and A. R. Scabra, “Pengaruh dosis perendaman ekstrak
daun cengkeh (Syzygium aromaticum) terhadap daya tetas telur ikan lele dumbo (Clarias
gariepinus),” Jurnal Perikanan Unram, vol. 11, no. 1, pp. 89–97, May 2021, doi:
10.29303/jp.v11i1.251.
[31] R. S. M. Chong, “Saprolegniasis,” Aquaculture Pathophysiology: Finfish Diseases: Volume I,
vol. 1, pp. 645–650, Jan. 2022, doi: 10.1016/B978-0-12-812211-2.00053-6.
[32] K. Chukanhom and K. Hatai, “Freshwater fungi isolated from eggs of the common carp
(Cyprinus carpio) in Thailand,” Mycoscience, vol. 45, no. 1, pp. 42–48, Feb. 2004, doi:
10.1007/s10267-003-0153-9.
12
[33] A. A. El-Shahir, D. A. El-Wakil, A. A. H. A. Latef, and N. H. Youssef, “Bioactive compounds

and antifungal activity of leaves and fruits methanolic extracts of Ziziphus spina-christi L.,”
Plants, vol. 11, no. 6, Mar. 2022, doi: 10.3390/plants11060746.
[34] B. Basir, and K. Kaharuddin, “Effectiveness of ketapang leaf extract (Terminal Capta L) in water
media on crude egg hatching,” International Journal on Advanced Science, Education, and
Religion, vol. 3, no. 1, pp. 33–39, Mar. 2020, doi: 10.33648/ijoaser.v3i1.44.
[35] A. Budiarti, D. N. Hidayati, and N. Aini, “Identifikasi awal potensi ekstrak etanol rimpang jahe
merah (Zingiber officinale Rosc. var. rubrum) sebagai obat osteoporosis.”
[36] D.G. Monroe, F.J. Secreto, and T.C. Spelsberg, “Overview of estrogen action in osteoblasts:
Role of the ligand, the receptor, and the co-regulators,” J Musculoskel Neuron Interact, vol. 3,
no. 4, pp. 357-362, 2003. PMID: 15758323.
[37] H. Andika, E. Dewantoro, and E. I. Raharjo, “Perendaman telur ikan gurame (Osphronemus
gouramy) dengan ekstrak meniran (Phyllanthus niruri L) sebagai anti jamur,” Jurnal Ruaya:
Jurnal Penelitian dan Kajian Ilmu Perikanan dan Kelautan, vol. 1, no. 1, Jan. 2013, doi:
10.29406/rya.v1i1.231.
[38] A. Baharudin, M. Bahrus Syakirin, and T. Yusufi Mardiana, “Pengaruh perendaman larutan teh
terhadap daya tetas telur ikan lele sangkuriang (Clarias gariepinus),” 2016.
[39] BSN, “Penetapan revisi 7 (tujuh) Standar Nasional Indonesia” 1. 14243_SK SNI no 148-KEP-
BSN-9-2014.
[40] B. R. Swistock, W. E. Sharpe, and T. McCarty, “Water quality concerns for ponds,” p. 6, 1998.
[41] I. Joseph, S. Joseph, B. Ignatius, G. S. Rao, K. S. Sobhana, D. Prema, and M. Varghese, “A pilot
study on culture of Asian seabass Lates calcarifer (Bloch) in open sea cage at Munambam,
Cochin coast, India,” Article in Indian Journal of Fisheries, 2010. [Online]. Available:
https://www.researchgate.net/publication/289202792
[42] L. J. Magnoni, O. Gonçalves, P. G. Cardoso, F. Silva-Brito, R. O. A. Ozório, M. Palma, and I.
Viegas, “Growth performance and quality indicators of European seabass (Dicentrarchus
labrax) fed diets including refined glycerol,” Aquaculture, vol. 569, May 2023, doi:
10.1016/j.aquaculture.2023.739377.
[43] R. B. Hasibuan, H. Irawan, and T. Yulianto, “The effect of temperature on hatched of seabass
egg,” Intek Akuakultur, vol. 2, no. 2, pp. 49–57, Oct. 2018, doi: 10.31629/intek.v2i2.539.
[44] M. Papadaki, M. Papadopoulou, I. Siggelaki, and C. C. Mylonas, “Egg and sperm production
and quality of sharpsnout sea bream (Diplodus puntazzo) in captivity,” Aquaculture, vol. 276,
no. 1–4, pp. 187–197, Apr. 2008, doi: 10.1016/j.aquaculture.2008.01.033.
13

IOP_Conf._Ser.__Earth_Environ._Sci._1289_012022

Uploaded by

Copyright:

Available Formats

IOP_Conf._Ser.__Earth_Environ._Sci._1289_012022

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

IOP_Conf._Ser.__Earth_Environ._Sci._1289_012022

Uploaded by

Copyright:

Available Formats

IOP Conference Series: Earth and Environmental Science

PAPER • OPEN ACCESS You may also like

- Effects of weight and storage duration of

This content was downloaded from IP address 114.122.21.20 on 31/12/2023 at 01:15

Leaf extract effectiveness of pecut kuda (Stachytarpheta

2.2 Research methodology

2.3 Research procedure

2.3.2 Container preparation

2.3.3 Preparation of barramundi eggs

2.3.4 Soaking test

2.3.5 Larvae rearing

2.4 Observation parameters

2.4.2 Fungal infection percentage

2.4.3 Larvae abnormality

2.4.4 Larval growth

2.4.5 Survival rate

2.4.6 Water quality parameters

2.5 Data analysis

Figure 1. Graph of egg hatchability in each treatment.

3.2 Fungus attack percentage

Figure 2. Fungus attack resistance level graph.

Figure 3. Saprolegnia sp which attacks barramundi fish eggs

3.3. Larvae abnormalities

Figure 4. Barramundi fish larvae

3.4. Growth length

Figure 5. Graph of specific growth rate for each treatment

3.5. Larvae mortalities

Table 2. Larvae mortality in each treatment.

100 84,98 88,19

Figure 6. Larvae survival rate.

3.6. Water quality parameters

Average water quality data

Table 4. Data on the measurement of water quality for rearing eggs

Average water quality data

[19] M. Kailasam, A. R. Thirunavukkarasu, A. G. Ponniah, S. Selvaraj, and P. Stalin, “Recent

[33] A. A. El-Shahir, D. A. El-Wakil, A. A. H. A. Latef, and N. H. Youssef, “Bioactive compounds

You might also like